Detection of snow mold fungi in wheat plants by means of polyacrylamide gel electrophoresis of ribosomal proteins.

Abstract

Ribosomes were isolated from four snow mold fungi (Pythium paddicum, P. iwayamai, Typhula incarnata and Fusarium nivale) and wheat seedlings, and their proteins were analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). No difference was found in electrophoretic patterns between P. paddicum and P. iwayamai. However, there were several differences among two Pythium species, T. incarnata, F. nivale and wheat. The ribosomes were isolated also from wheat seedlings infected with P. paddicum, T. incarnata and F. nivale individually, and their proteins were analyzed by SDS-PAGE. The distinctive fungal ribosomal protein bands were detected in all patterns obtained from the infected wheat plants. The possibility of the ribosomal protein-based assay to detect snow mold fungi in wheat plants is suggested.

Immunological techniques used with fungal plant pathogens: aspects of antigens, antibodies and assays for diagnosis

“…Ribosomal proteins and enzymes have also been examined as immunogens. Ribosomal epitopes seem to be specific to the fungus under examination for instance with Fusarium moniliforme (Marshall & Partridge, 1981); snow mould fungi (Takenaka, 1988(Takenaka, , 1992. The same applies to enzymes such as invertase: Phytophthora megasperma f.sp.…”

“…Background reactions can be reduced or eliminated by the addition of different blocking agents which saturate non-specific binding areas. These include for fungi (not only in ELISA) skimmed milk (Gleason et al, 1987;Wright & Morton, 1989), gelatine (El-Nashaar, Moore & George, 1986; Fuhrmann, Roquebert, Van Hoegaerden & Strossberg, 1989;Suske & Acker, 1989), bovine serum albumin (BSA) Mohan, 1988Mohan, , 1989aTakenaka, 1988;Burdsall et al, 1990;Phelps et al, 1990;Priestley, 1990), albumin from chicken protein (Musgrave & Fletcher, 1986), and serum of non-immunised animals (Priestley, 1990).…”

“…Although successful with viruses and bacteria. this method is not good for the diagnosis of fungi (Takenaka, 1988(Takenaka, , 1992. It is useful for analysis of antigen structures or the specificity of antigenantibody-interactions (Benhamou.…”

Evaluation of a polyclonal antibody immunoassay for detection and quantification of Macrophomina phaseolina

“…These suspensions were stored at ¹20ЊC and used as soluble protein and cell wall antigens, respectively. Ribosomes were obtained from mycelial homogenate of M. phaseolina grown for 6 days in SM (Takenaka & Yoshino, 1987). The homogenate was ultracentrifuged through a sucrose pad and treated with bovine pancreatic ribonuclease-A (Sigma, St Louis, MO, 1 mg mL ¹1 ) at 4ЊC for 30 min.…”